Interaccion de promastigotes de Leishmania donovani con celulas de exudado peritonial de ratones mediante la influencia de la quimotripsina

Arcay, Lucila; Bruzual, Elizabeth

doi:10.1590/S0036-46651986000300002

Resúmenes

Se hace un estudio de la interacción de promastigotes de Leishmania donovani con células de exudado peritoneal de ratón (c e p) mediante la influencia de la quimotripsina. La adhesión de los promastigotes a las c e p fue terminal y marginal, y en observaciones hechas a partir de los 10 minutos de enfrentamiento, esta adhesión fue nula hasta los 30 minutos en el grupo tratado, y sólo a las das horas hubo un pequeño incremento (2.4%) con respecto al control. Se observa marcada disminución en todos los parámetro medidos, tales como enlace, penetración, multiplicación intracelular, división de formas flageladas, en el grupo tratado. La quimotripsina favorece la formación de formas intermedias flageladas, heciendose el parásito piriforme y esférico, apareciendo una forma aberrante de extremo anterior cilindrico que semeja a una forma coanoflagelada. Se sospecha que la enzima reduce efectivamente fragmentos proteicos o péptidos, los cuales pueden haber sido tan pequeños como para esconder a otros ligandos relacionados con la adhesión macrófago-parásito.

Calazar; Leishmania donovani; Peritonio; Ratos

The interaction of promastigotes of Leishmania donovani with mouse peritoneal exudate cells (cep) under the influence of Chymotripsine was studied. The promastigote adhered to the cep terminally and marginally. At 10-30 minutes postchallange, adhesion was absent in the treated group, and only after two hours was there any adhesión (24% in comparison with the controls). The experimental group was markedly deficient in all the parameters of activity measured: attachment, penetration, intracellular multiplication and division of flagellates forms in comparison to controls. The Chymotripsine favored the development of intermediate flagellates forms, these being spheroid or pyriform, with example of an aberrant form having the anterior end cylindrical similar to a choanoflagellate. It is suspected that the Chymotripsine effectively reduces protein or peptide fragments; these may have been so small as to conceal other ligands associated with macrophage-parasite adhesion.

ARTIGOS ORIGINAIS

Interaccion de promastigotes de Leishmania donovani con celulas de exudado peritonial de ratones mediante la influencia de la quimotripsina

Interaction of Leishmania donovani promastigotes with mouse peritoneal exudate cells under the influence of the Chymotripsine

Lucila Arcay^I; Elizabeth Bruzual^II

^IInstituto de Zoologia Tropical, Universidad Central de Venezuela, Caracas, Venezuela

^IIEscuela de Medicina "José Maria Vargas", Universidad Central Caracas, Venezuela

RESUMEN

Se hace un estudio de la interacción de promastigotes de Leishmania donovani con células de exudado peritoneal de ratón (c e p) mediante la influencia de la quimotripsina. La adhesión de los promastigotes a las c e p fue terminal y marginal, y en observaciones hechas a partir de los 10 minutos de enfrentamiento, esta adhesión fue nula hasta los 30 minutos en el grupo tratado, y sólo a las das horas hubo un pequeño incremento (2.4%) con respecto al control. Se observa marcada disminución en todos los parámetro medidos, tales como enlace, penetración, multiplicación intracelular, división de formas flageladas, en el grupo tratado. La quimotripsina favorece la formación de formas intermedias flageladas, heciendose el parásito piriforme y esférico, apareciendo una forma aberrante de extremo anterior cilindrico que semeja a una forma coanoflagelada. Se sospecha que la enzima reduce efectivamente fragmentos proteicos o péptidos, los cuales pueden haber sido tan pequeños como para esconder a otros ligandos relacionados con la adhesión macrófago-parásito.

Unitermos: Calazar Leishmania donovani Peritonio Ratos.

SUMMARY

The interaction of promastigotes of Leishmania donovani with mouse peritoneal exudate cells (cep) under the influence of Chymotripsine was studied. The promastigote adhered to the cep terminally and marginally. At 10-30 minutes postchallange, adhesion was absent in the treated group, and only after two hours was there any adhesión (24% in comparison with the controls). The experimental group was markedly deficient in all the parameters of activity measured: attachment, penetration, intracellular multiplication and division of flagellates forms in comparison to controls. The Chymotripsine favored the development of intermediate flagellates forms, these being spheroid or pyriform, with example of an aberrant form having the anterior end cylindrical similar to a choanoflagellate. It is suspected that the Chymotripsine effectively reduces protein or peptide fragments; these may have been so small as to conceal other ligands associated with macrophage-parasite adhesion.

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AGRADECIMIENTO

Al histotecnólogo del Instituto de Zoologia Tropical de la Universidad Central de Venezuela (UCV) por su asistencia técnica y dibujo lineal de los gráficos. Al Consejo de Desarollo Cientifico y Humanístico de Ir UCV por la subvención del Proyecto.

Recebido para publicação em 17/4/1985.

1. ABAKAROVA, E. G. & AKINSHINA, G. T. Interaction between macrophages of various origin and Toxoplasma gondii under condtions "in vitro". Folia Parasit., 22: 195-200, 1975.
2. ALEXANDER, J. & VICKERMAN, K. Fusion of host cell secondary lysosomes with the parasitophorous vacuoles of Leishmania mexicana infect macrophages. J. Protozoal., 22: 502-508, 1975.
3. ARCAY, L. Influencia de las hormonas sexuales sobre la infección experimental producida por una cepa de Leishmania mexicana amazonensis de Venezuela. Rev. lat. amer. Microbiol., 27: 195-207, 1985.
4. ARCAY -DE-PER AZA, L.; OCHOA, N. & ERCOLI, N. Nuevas lesiones genitales ocasionadas por Leishmania mexicana amazonensis. Acta cient. venez., 29: 34-41, 1978.
5. BASTARDO, T.; ARCAY, L. & TEJERO, F. Interaction of avirulent Leishmania, species with rat peritoneal macrophages. Mem. Inst. Osw. Cruz, 18: 21-26, 1983.
6. BRUZUAL, E. & ARCAY, L. Visceralizacion generalizada de Leishmania mexicana amazonensis en hamsters y ratones, bajo la influencia de la quimotripsina, 1985. (en prensa)
7. BIANCO, C; GRIFFEN, F. N. & SIL VERSTEIN, S. C. Studies on macrophages complement receptor. Alteration of receptor function upon macrophage activation. J. exp. Med., 141: 1278-1290, 1975.
8. BRAY, R. S. Leishmania mexicana mexicana: attachment and uptake of promastigotes and macrophages in vitro. J. Protozool., 30: 314-322, 1983a.
9. BRAY, R. S. Leishmania: Chemotaxic responses of promastigotes and macrophages in vitro. J. Protozool., 30: 322-329, 1983b.
10. COUTINHO, E. & COELHO, M. V. Experimental Cutaneous Leishmaniasis. II. The pathology of Leishma- niasis by L. mexicana. Rev. Inst. Med. trop. S. Paulo, 7: 145-155, 1965.
11. CUATRECASAS, P. & HOLLEMBERG, M. D. Membrane receptors and hormone action. Advanc. Protein Chem., 30: 251-451, 1976.
12. CHANG, K. P. Endocytosis of Leishmania-infected macrophages, fluorometry of pinocytic rate, lysosome phagozome fusion and intralysosomal pH. In: VAN DEN BOSSCHE, H., ed. The host invader Interplay Jansen Research Foundation. Amsterdam, Elsevier/ Nfich-Holland Biomedical Press, 1980. p. 231-234.
13. CHANG, K. F. & DWYER, D. M. Leishmania donovani hamster peritoneal macrophage interactions in vitro. J. Parasit., 61 (Suppl.): 71, 1975.
14. GIANNINI, M. S. H. ; D'ALESANDRO, P. A.; GARCIA, C. R. & SARAIVA, E. M. B. Failure of hamster macrophages to discriminate between infective and non infective promastigotes of Leishmania donovani during attachment in vitro. Infect. Immun. 34: .629-632, 1981.
15. GUIMARÃES, F. N. Leishmaniose experimental. Visceralizagão de Leishmania brasiliensis Vianna, 1911 em hamsters (Cricetus auratus Waterhouse). Brasil méd., 61: Separatim, 47-48, 1947.
16. GUIMARÃES, F. N. Leishmaniose experimental. II. Comportamento de L. brasil lensis Vianna 1911 em hamsters (Cricetus (Mesocricetus) auratus Waterhouse). Hospital (Rio de J.), 40: 25-45, 1951a.
17. GUIMARÃES, P. N. Leishmaniose experimental. III. a) Comportamento de L. brasiliensis em camundongo (Mus musculas albina), b) Infecções ligeiras em cotton-rats (Sigmodon hispidus). c) Animais refractorios. Hospital (Rio de J.), 40: 153-162, 1951b.
18. GUIMARÃES, F. N. Leishmaniose experimental. IV. Reprodução em hamsters (Cricetus auratus) de uma leishmaniose cutânea núdulo-tumoral oriunda da Amazonia. Hospital (Rio de J.), 40: 665-676, 1951c.
19. GUIMARÃES, F. N, Leishmaniose experimental. V. Reprodução em camundongos (Mus musculus albina) de uma leishmaniose cutânea nódulo-tumoral (histiocitoma leishmânico), ocorrendo na Amazonia. Hospital (Rio de J.), 40: 919-931, 1951d.
20. HERMAN, R. Cytophilic and opsonic antibodies in visceral leishmaniasis in mice. Infect. Immun., 28: 585-593, 1980.
21. HERNANDEZ, A. G.; INFANTE, R. B.; LA RIVA, G. de V. O.; DAWIDOWICZ, K. & CONVIT, J. The surface membrane of Leishmania. II. Energy dependent agglutination of Leishmania brasiliensis by concanavalin A. Acta cient. venez., 28: 380-384, 1977.
22. KEUSCH, G. T. Specific membrane receptors: pathogenetic and therapeutic implications in infections diseases. Rev. infect. Dis., 1: 517-529, 1979.
23. KEUSCH, G. T. The role of bacterial adherence in infection In: MAGNO, G.; COTRAN, R. S. & KAUFMAN, N., ed. Current topics In Inflammation Baltimore, William & Wilkins, 1982. p. 94-113.
24. LEWIS, D. H. & PETERS, W. The resistance of intracellular Leishmania parasites to digestion by lisosomal enzimes. Ann. trop. Med. Parasit., 71: 295-312, 1977.
25. MEDINA, R. & ROMERO, J. Leishmania pifanol n. sp., el agente causal de la leishmaniasis tegumentaria difusa. Arch. venez. Med. trop., 4: 349-353, 1962.
26. OCHOA DE MACHADO, N. & ARCAY-DE-PERAZA, L. Diseminación de Leishmania mexicana amazonensis mediante la inoculación intracardiaca, tajo la influencia de la acción de la mucina gástrica. Acta cient. venez., 29: 119-125, 1978.
27. SILVERSTEIN, S. C. Endocytic uptake of particles by mononuclear phagocytes and the penetration of obligate intracellular parasites. Amer. J. trop. Med. Hyg., 26: 161-169, 1977.
28. SCHNUR. L. P.; ZUCKERMAN, A. & MONTILLO, B. Dissemination of leishmanias to the organs of Syriam hamsters following intrasplenic inoculation of promastigotes. Exp. Parasit., 34: 432-447, 1973.
29. STAUBERT, L. A. Leishmaniasis in hamsters. In: Some-physiological aspects and consequences of parasitism New Brunswick, Rutgers Univ. Press, 1955. p. 76-90.
30. SUTTER, E. Multiplication of tubercle bactli within mononuclear phagocytes in tissue culture derived from normal animal vaccined with BCG. J. exp. Med., 97: 235-245, 1953.
31. WERB, Z. Phagocytic cells: Chemotaxis and effects funtion of macrophages and granulocytes. In: WITZ, I. P. & HAND A, M. O., ed. Contemporary topics In immunoblology New York, Plenum, 1981. p. 109-123.
32. WONDE, M. M. T. & LAMY, L. Differences de comportment de Leishmania donovanl dans les macrophages de souris, de rat et hamster in vivo et in vitro. C.R. Acad. Sci. (Paris) (Serie D) 265: 810-813, 1967.
33. WYLER, D. & SUZUKI, R. In vitro parasite-monocyte interaction in human leishmaniasis: effect of enzime treatments on attachment. Infect. Immun., 42: 356-361, 1983.
34. ZENIAN, A.; ROWLES, P. & GINGELL, D. Scanning Electron-microscopic study of the uptake of Leishmania parasites by macrophages. J. Cell. Sci., 39: 187-199, 1979.
35. ZUCKERMAN, A. Current status of the immunology of blood and tissue protozoa. I. Leishmania. Exp. Parasit., 38: 370-400, 1975.

Fechas de Publicación

Publicación en esta colección
16 Oct 2012
Fecha del número
Jun 1986

This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License.

[1] 1. ABAKAROVA, E. G. & AKINSHINA, G. T. Interaction between macrophages of various origin and Toxoplasma gondii under condtions "in vitro". Folia Parasit., 22: 195-200, 1975.

[2] 2. ALEXANDER, J. & VICKERMAN, K. Fusion of host cell secondary lysosomes with the parasitophorous vacuoles of Leishmania mexicana infect macrophages. J. Protozoal., 22: 502-508, 1975.

[3] 3. ARCAY, L. Influencia de las hormonas sexuales sobre la infección experimental producida por una cepa de Leishmania mexicana amazonensis de Venezuela. Rev. lat. amer. Microbiol., 27: 195-207, 1985.

[4] 4. ARCAY -DE-PER AZA, L.; OCHOA, N. & ERCOLI, N. Nuevas lesiones genitales ocasionadas por Leishmania mexicana amazonensis. Acta cient. venez., 29: 34-41, 1978.

[5] 5. BASTARDO, T.; ARCAY, L. & TEJERO, F. Interaction of avirulent Leishmania, species with rat peritoneal macrophages. Mem. Inst. Osw. Cruz, 18: 21-26, 1983.

[6] 6. BRUZUAL, E. & ARCAY, L. Visceralizacion generalizada de Leishmania mexicana amazonensis en hamsters y ratones, bajo la influencia de la quimotripsina, 1985. (en prensa)

[7] 7. BIANCO, C; GRIFFEN, F. N. & SIL VERSTEIN, S. C. Studies on macrophages complement receptor. Alteration of receptor function upon macrophage activation. J. exp. Med., 141: 1278-1290, 1975.

[8] 8. BRAY, R. S. Leishmania mexicana mexicana: attachment and uptake of promastigotes and macrophages in vitro. J. Protozool., 30: 314-322, 1983a.

[9] 9. BRAY, R. S. Leishmania: Chemotaxic responses of promastigotes and macrophages in vitro. J. Protozool., 30: 322-329, 1983b.

[10] 10. COUTINHO, E. & COELHO, M. V. Experimental Cutaneous Leishmaniasis. II. The pathology of Leishma- niasis by L. mexicana. Rev. Inst. Med. trop. S. Paulo, 7: 145-155, 1965.

[11] 11. CUATRECASAS, P. & HOLLEMBERG, M. D. Membrane receptors and hormone action. Advanc. Protein Chem., 30: 251-451, 1976.

[12] 12. CHANG, K. P. Endocytosis of Leishmania-infected macrophages, fluorometry of pinocytic rate, lysosome phagozome fusion and intralysosomal pH. In: VAN DEN BOSSCHE, H., ed. The host invader Interplay Jansen Research Foundation. Amsterdam, Elsevier/ Nfich-Holland Biomedical Press, 1980. p. 231-234.

[13] 13. CHANG, K. F. & DWYER, D. M. Leishmania donovani hamster peritoneal macrophage interactions in vitro. J. Parasit., 61 (Suppl.): 71, 1975.

[14] 14. GIANNINI, M. S. H. ; D'ALESANDRO, P. A.; GARCIA, C. R. & SARAIVA, E. M. B. Failure of hamster macrophages to discriminate between infective and non infective promastigotes of Leishmania donovani during attachment in vitro. Infect. Immun. 34: .629-632, 1981.

[15] 15. GUIMARÃES, F. N. Leishmaniose experimental. Visceralizagão de Leishmania brasiliensis Vianna, 1911 em hamsters (Cricetus auratus Waterhouse). Brasil méd., 61: Separatim, 47-48, 1947.

[16] 16. GUIMARÃES, F. N. Leishmaniose experimental. II. Comportamento de L. brasil lensis Vianna 1911 em hamsters (Cricetus (Mesocricetus) auratus Waterhouse). Hospital (Rio de J.), 40: 25-45, 1951a.

[17] 17. GUIMARÃES, P. N. Leishmaniose experimental. III. a) Comportamento de L. brasiliensis em camundongo (Mus musculas albina), b) Infecções ligeiras em cotton-rats (Sigmodon hispidus). c) Animais refractorios. Hospital (Rio de J.), 40: 153-162, 1951b.

[18] 18. GUIMARÃES, F. N. Leishmaniose experimental. IV. Reprodução em hamsters (Cricetus auratus) de uma leishmaniose cutânea núdulo-tumoral oriunda da Amazonia. Hospital (Rio de J.), 40: 665-676, 1951c.

[19] 19. GUIMARÃES, F. N, Leishmaniose experimental. V. Reprodução em camundongos (Mus musculus albina) de uma leishmaniose cutânea nódulo-tumoral (histiocitoma leishmânico), ocorrendo na Amazonia. Hospital (Rio de J.), 40: 919-931, 1951d.

[20] 20. HERMAN, R. Cytophilic and opsonic antibodies in visceral leishmaniasis in mice. Infect. Immun., 28: 585-593, 1980.

[21] 21. HERNANDEZ, A. G.; INFANTE, R. B.; LA RIVA, G. de V. O.; DAWIDOWICZ, K. & CONVIT, J. The surface membrane of Leishmania. II. Energy dependent agglutination of Leishmania brasiliensis by concanavalin A. Acta cient. venez., 28: 380-384, 1977.

[22] 22. KEUSCH, G. T. Specific membrane receptors: pathogenetic and therapeutic implications in infections diseases. Rev. infect. Dis., 1: 517-529, 1979.

[23] 23. KEUSCH, G. T. The role of bacterial adherence in infection In: MAGNO, G.; COTRAN, R. S. & KAUFMAN, N., ed. Current topics In Inflammation Baltimore, William & Wilkins, 1982. p. 94-113.

[24] 24. LEWIS, D. H. & PETERS, W. The resistance of intracellular Leishmania parasites to digestion by lisosomal enzimes. Ann. trop. Med. Parasit., 71: 295-312, 1977.

[25] 25. MEDINA, R. & ROMERO, J. Leishmania pifanol n. sp., el agente causal de la leishmaniasis tegumentaria difusa. Arch. venez. Med. trop., 4: 349-353, 1962.

[26] 26. OCHOA DE MACHADO, N. & ARCAY-DE-PERAZA, L. Diseminación de Leishmania mexicana amazonensis mediante la inoculación intracardiaca, tajo la influencia de la acción de la mucina gástrica. Acta cient. venez., 29: 119-125, 1978.

[27] 27. SILVERSTEIN, S. C. Endocytic uptake of particles by mononuclear phagocytes and the penetration of obligate intracellular parasites. Amer. J. trop. Med. Hyg., 26: 161-169, 1977.

[28] 28. SCHNUR. L. P.; ZUCKERMAN, A. & MONTILLO, B. Dissemination of leishmanias to the organs of Syriam hamsters following intrasplenic inoculation of promastigotes. Exp. Parasit., 34: 432-447, 1973.

[29] 29. STAUBERT, L. A. Leishmaniasis in hamsters. In: Some-physiological aspects and consequences of parasitism New Brunswick, Rutgers Univ. Press, 1955. p. 76-90.

[30] 30. SUTTER, E. Multiplication of tubercle bactli within mononuclear phagocytes in tissue culture derived from normal animal vaccined with BCG. J. exp. Med., 97: 235-245, 1953.

[31] 31. WERB, Z. Phagocytic cells: Chemotaxis and effects funtion of macrophages and granulocytes. In: WITZ, I. P. & HAND A, M. O., ed. Contemporary topics In immunoblology New York, Plenum, 1981. p. 109-123.

[32] 32. WONDE, M. M. T. & LAMY, L. Differences de comportment de Leishmania donovanl dans les macrophages de souris, de rat et hamster in vivo et in vitro. C.R. Acad. Sci. (Paris) (Serie D) 265: 810-813, 1967.

[33] 33. WYLER, D. & SUZUKI, R. In vitro parasite-monocyte interaction in human leishmaniasis: effect of enzime treatments on attachment. Infect. Immun., 42: 356-361, 1983.

[34] 34. ZENIAN, A.; ROWLES, P. & GINGELL, D. Scanning Electron-microscopic study of the uptake of Leishmania parasites by macrophages. J. Cell. Sci., 39: 187-199, 1979.

[35] 35. ZUCKERMAN, A. Current status of the immunology of blood and tissue protozoa. I. Leishmania. Exp. Parasit., 38: 370-400, 1975.

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Interaccion de promastigotes de Leishmania donovani con celulas de exudado peritonial de ratones mediante la influencia de la quimotripsina

Interaction of Leishmania donovani promastigotes with mouse peritoneal exudate cells under the influence of the Chymotripsine

Resúmenes

Fechas de Publicación